W e work primarily with mouse models where we can examine T cell responses in vivo at a high resolution. Experiments in human cells and monitoring immunity in non-human primates are also used in the lab. We use transgenic and knockout mice of various flavors routinely - and generate some of our own. The mechanisms we study are relevant to a wide variety of patho-physiological contexts. So, depending on the specific experimental question, we may use the following mouse models available in the lab ::
- Psoriasis / Dermatitis
- Hyperacute rejection
- Solid Graft rejection
- Influenza PR8
- Pneumonia Virus of Mice
- Yellow Fever - YF17D
- HPV (Gardasil)
- Pertussis (whole cell and acellular)
- HCV - experimental
- Melanoma (B16)
The lab uses a wide variety of cellular and molecular techniques as required by particular experiments. On any given week we may be doing any or all of these in the lab...
- Isolation of primary T, B and Dendritic cells, magnetic bead purifications, MACS etc.
- Flow Cytometry & Cell sorting
- Overexpression/Knockdown of genes in primary lymphocytes using Amaxa nucleofaction, Retroviral transduction or Electroporation
- Retrogenic mice - retroviral transduction of primary Bone Marrow stem cells followed by generation of chimeric mice (primarily to clone & express specific T cell receptors)
- ELISAs, Luminex, Bead array & other cytokine measurements
- TCR repertoire analysis, MHC tetramers, expression in insect cells
- Culture, growth and infection with parasites, viruses
- Cell culture and in vitro assays for T or DC function
- In-house cell fusion and generation of monoclonal antibodies
- RNAseq, single-cell RNAseq & CITEseq
- single-cell ATACseq & ASAPseq
- Standard molecular biology & biochemistry techniques.
eagents from the lab:
- Plasmids generated by us are available from the Addgene repository: https://www.addgene.org/Nevil_Singh/
- Contact the Lab directly for any mouse strains.